The present invention relates to a heterocyclic ring condensed benzothiazine compound useful as a medicament, a medicament containing the same and processes for producing the same. More specifically, it relates to a novel heterocyclic ring condensed benzothiazine compound useful as a medicament for diseases against which the effect of inhibiting the binding of IgE receptor xcex3 and a tyrosine kinase of 72 kDa is effective.
The bronchial asthma and the atopic diseases in human beings appear in consequence of highly intriacate vital reactions. It is suspected that most of these conditions are caused because various chemical mediators liberated from mast cells and basophils, as triggered by antigen-antibody reactions, induce vital disturbances as by contracting such smooth muscles as bronchial muscles and vessels of the pulmonary circulation or enhancing permeability of blood vessels.
As the chemical mediators liberated from mast cells and basophils, histamine, leukotrienes, prostaglandins, TNF, etc. have been known. It is well known that histamine, among other substances mentioned above, is the most significant chemical mediator for the allergic rhinitis and the urticaria in human beings. The leucotrienes comprise leucotrienes B4, C4, and D4 and the relation thereof with the asthmatic convulsion has been attracting attention.
Heretofore, the development of medicines for the prevention, alleviation, or elimination of the symptoms of allergic diseases has been aimed at repressing the creation and liberation of such chemical mediators or antagonizing the effects thereof.
Sodium cromoglycate (Intal(trademark)) having been marketed since 1969 is a typical example of these drugs.
However, the conventional antiallergic agents typified by Intal(trademark) show difference in the chemical mediator liberation inhibitory concentration between in vitro and in vivo. Moreover, sensitivities to these drugs widely vary from patient to patient and their action mechanisms still remain unknown in many points.
Mast cells and basophils closely relating to allergic diseases have a highly affinitive receptor, Fc xcex5 RI, for the IgE antibody on the cell membrane thereof. IgE antibody""s binding to this receptor forms a cross-linkage with the. corresponding polyvalent antigen, the intracellular signal transmission mechanism is activated. Then histamine is liberated or leukotrienes and prostaglandins are formed and liberated, thus inducing the onset of the so-called allergic symptoms. It is furthermore considered that the cytokines such as TNF and interleukins thus produced interact with other cells and thus make the diseases chronic.
Under these circumstances, the present inventors have paid their attention to the activation of a non-receptor type tyrosine kinase located at the early stage in the activation of the intracellular signal transmission mechanism upon liberation of chemical mediators from mast cells or basophils. It is known that this tyrosine kinase is activated when it binds to the phosphorylated tyrosine activation motif (TAM) region in the IgE receptor xcex3 chain. By inhibiting this binding to thereby inhibit the activation of the tyrosine kinase of 72 kDa, the activation of the intracellular signal transmission mechanism depending on the IgE antibody in mast cells or basophils can be inhibited. As a result, also the liberation of the above chemical mediators can be inhibited. The present inventors have found out that desired objects can be achieved by using heterocyclic ring condensed benzothiazine compounds represented by the following formula (I), thus completing the present invention.
WO97/33871 discloses a phenothiazine compound as a prophylactic or therapeutic agent for allergic disease of the similar mechanism, but does not disclose nor suggest a compound having a benzene ring containing a hetero atom.
An object of the invention is to provide a novel acridone compound which is effective for prevention or remedy of asthma, allergic coryza, atopic dermatitis, hives, hay fever, gastrointestinal allergy, food allergy and the like, and a pharmacologically acceptable salt thereof, and another object thereof is to provide a medicine containing, as an active ingredient, the compound, a hydrate thereof or a pharmacologically acceptable salt thereof.
That is, the invention is a heterocyclic ring condensed benzothiazine compound represented by the following formula (I), its pharmacologically acceptable salt or hydrates thereof. Formula (I) 
Wherein, the ring Het represents an unsaturated heterocyclic ring that may be substituted with from 1 to 3 substituents;
R1 and R2 are the same as or different from each other, and each represents halogen atom, a lower alkyl group that may be substituted with halogen, a lower alkoxy group that may be substituted with halogen, a lower alkoxy lower alkyl group, cyano group, carbamoyl group that may have a substituent or a carboxyl group that may have a protective group, and R1 and R2 may form a ring along with the carbon atom to which they are bound. The ring may contain oxygen atom, sulfur atom or nitrogen atom, and may have a substituent;
D represents a lower alkylene chain that may have a substituent, a lower alkenylene chain that may have a substituent, a lower alkynylene chain that may have a substituent or 
(in the formula, m and 1 represent an integer of from 0 to 6, and the ring A represents a hydrocarbon ring that may have a substituent or a heterocyclic ring that may have a substituent)
Q represents carbamoyl group that may have a substituent, an acyl group, an acyl lower alkyl group, a carboxyl group that may have a protective group, a heteroaryl group that may have a substituent or a formula xe2x80x94NR20R21. In the formula xe2x80x94NR20R21, R20 and R21 are the same as or different from each other, and each represents hydrogen atom, a lower alkyl group that may be substituted with halogen, a lower alkoxy group that may be substituted with halogen, a lower alkyl group that is substituted with a hydroxyl group, a lower alkoxy lower alkyl group, an aryl group that may have a substituent, an aryl lower alkyl group that may have a substituent, a heteroaryl group that may have a substituent, a heteroaryl lower alkyl group that may have a substituent, an aryloxy group that may have a substituent, an aryl lower alkoxy group that may have a substituent, a heteroaryloxy group that may have a substituent, a heteroaryl lower alkoxy group that may have a substituent, a carboxyalkyl group that may have a protective group, an acyl group, an acyl lower alkyl group that may have a substituent, an acylamino group that may have a substituent, an acylamino lower alkyl group that may have a substituent, a carbamoyl lower alkyl group that may have a substituent, an amino lower alkyl group that may have a substituent, a cyano lower alkyl group, an acyl lower alkyl group, a lower cycloalkyl group, a lower cycloalkyl lower alkyl group or an amidino group that may be substituted with a lower alkyl group. Alternatively, R20 and R21 may form a 3- to 8-membered ring along with the nitrogen atom to which they are bound, and the ring may have, as a component constituting the ring, in addition to the carbon atoms at least one selected from the group consisting of a nitrogen atom, a sulfur atom, an oxygen atom and a formula xe2x80x94NR22, and may have a substituent.
In the formula xe2x80x94NR22, R22 represents hydrogen atom, a lower alkyl group that may be substituted with halogen, an acyl group, an acyl lower alkyl group that may have a substituent, an aryl group that may have a substituent, a heteroaryl group that may have a substituent, an arylalkyl group that may have a substituent, a heteroarylalkyl group that may have a substituent or a formula
xe2x80x94S(O)sxe2x80x94(Y)uxe2x80x94R23.
In the formula xe2x80x94S(O)sxe2x80x94(Y)uxe2x80x94R23, R23 represents hydrogen atom, a lower alkyl group that may be substituted with halogen or an aryl group that may have a substituent, Y represents a methylene chain, s represents an integer of from 0 to 2, and u represents 0 or 1); and
x represents an integer of from 0 to 2.
The present invention provides a tyrosine kinase inhibitor, a preventive or therapeutic agent for disease, against which a tyrosine kinase inhibiting action is effective, an antiallergenic drug, a preventive or therapeutic agent for disease, against which an antiallergenic action is effective, or a preventive or therapeutic agent for asthma, allergic coryza, atopic dermatitis, hay fever, allergic conjunctivitis and food allergy, which comprises the heterocyclic ring condensed benzothiazine compound represented by the above formula (I) its pharmacologically acceptable salt or hydrates thereof as an active ingredient.
The present invention provides a method and a use for inhibiting tyrosine kinase, prevention or remedy of disease, against which a tyrosine kinase inhibiting action is effective, antiallergy, and disease, against which an antiallergenic action is effective, and prevention and remedy of asthma, allergic coryza, atopic dermatitis, hay fever, allergic conjunctivitis and food allergy, by administering a pharmacologically or clinically effective amount of the heterocyclic ring condensed benzothiazine compound represented by the above formula (I), its pharmacologically acceptable salt or hydrates thereof, to a patient infected with disease, against which tyrosine kinase inhibiting action is effective.
The present invention provides a medical composition containing a pharmacologically or clinically effective amount of the heterocylic ring condensed benzothiazine compound represented by the formula (I), its pharmacologically acceptable salt or hydrates thereof, and a pharmacologically acceptable carrier.
In the formula (I), the unsaturated hetero ring means a monocyclic heterocyclic ring, such as thiophene, furan, pyridine, pyrimidine, pyrazine, pyridazine and the like, or a bicyclic heterocyclic ring, such as quinoline, isoquinoline, naphthyridine.
The halogen atom found in the definition of R1 and R2 means fluorine, chlorine, bromine and iodine.
The lower alkyl group in the lower alkyl group that may be substituted with halogen found in the definition of R1, R2 R22 and R23 means a linear or branched alkyl group having from 1 to 6 carbon atoms. Examples thereof include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, n-pentyl group, 1,2-dimethylpropyl group, 1,1-dimethylpropyl group, 2,2-dimethylpropyl group, 2-ethylpropyl group, n-hexyl group, 1,2-dimethylbutyl group, 2,3-dimethylbutyl group, 1,3-dimethylbutyl group, 1-ethyl-2-methylpropyl group, 1-methyl-2-ethylpropyl group and the like.
The language xe2x80x9cmay be substituted with halogenxe2x80x9d herein means that the above alkyl groups may be substituted with from 1 to 3 halogen atoms, such as fluorine, chlorine, bromine and iodine. That is, trifluoromethyl group, dibromoethyl group and the like are included in the lower alkyl group that may be substituted with halogen in the formula (I).
The cycloalkyl group of the cycloalkyl group that may have a substituent found in the definition of R20 and R21 means those having from 3 to 8 carbon atoms, such as cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group, cycloheptyl group and cyclooctyl group.
The cycloalkylalkyl group found in the definition of R20 and R21 means those wherein the above cycloalkyl group is attached to any carbon atom of the above lower alkyl group.
The lower alkoxy group of the lower alkoxy group that may be substituted with halogen found in the definition of R1, R2 R20 and R21 means a linear or branched alkoxy group having from 1 to 6 carbon atoms. Examples thereof include methoxy group, ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, sec-butoxy group, tert-butoxy group, n-pentyloxy group,. 1,2-dimethylpropyloxy group, 1,1-dimethylpropyloxy group, 2,2-dimethylpropyloxy group, 2-ethylpropyloxy group, n-hexyloxy group, 1,2-dimethylbutyloxy group, 2,3-dimethylbutyloxy group, 1,3-dimethylbutyloxy group, 1-ethyl-2-methylpropyloxy group, 1-methyl-2-ethylpropyloxy group and the like.
The term xe2x80x9cmay be substituted with halogenxe2x80x9d herein means that the above alkoxy group may be substituted with from 1 to 3 halogen atoms, such as fluorine, chlorine, bromine, iodine. That is, trifluoromethoxy group, dibromoethoxy group and the like are also included in the lower alkoxy group that may be substituted with halogen in the present invention.
The acyl group found in the definition of R20, R21 and R22 means a group derived from an aliphatic saturated monocarboxylic acid, such as acetyl group, propyonyl group, butyryl group, valeryl group, isovaleryl group, pivaloyl group and the like, a group derived from an aliphatic unsaturated carboxylic acid, such as acryloyl group, propyoloyl group, methacryloyl group, crotonoyl group, isocrotonoyl group and the like, a group derived from a carbocyclic carboxylic acid, such as benzoyl group, naphthoyl group, toluoyl group, hydroatropoyl group, atropoyl group, cinnamoyl group and the like, a group derived from a heterocyclic carboxylic acid, such as furoyl group, thenoyl group, nicotinoyl group, isonicotinoyl group and the like, a group derived from a hydroxycarboxylic acid or an alkoxycarboxylic acid, such as glycoloyl group, lactoyl group, glyceloyl group, tropoyl group, benzyloyl group, salicyloyl group, anisoyl group, vanilloyl group, piperonyloyl group, galloyl group and the like, and a group derived from various amino acids.
The acylalkyl group found in the definition of R20, R21and R22 means those wherein the acyl group defined above is attached to any carbon atom of the lower alkyl group defined above. Examples thereof include acetylmethyl group, propyonylmethyl group, benzoylethyl group, naphthoylpropyl group, cinnamoylpropyl group, saliciloylbutyl group, nicotinoylpentyl group, glyceloylhexyl group and the like, but it is not limited thereto.
The alkoxyalkyl group found in the definition of R1, R2, R20 and R21 means those wherein the lower alkoxy group defined above is attached to any carbon atom of the lower alkyl group defined above. It means methoxymethyl group, ethoxymethyl group, ethoxyethyl group, 2-ethoxypropyl group and the like, but is not limited thereto.
The cyanoalkyl group found in the definition of R20, R21, R22 and R23 means those wherein the cyano group is attached to any carbon atom of the lower alkyl group defined above. It means cyano methyl group, 1-cyanoethyl group, 2-cyanoethyl group, 1-cyanopropyl group, 2-cyanopropyl group and the like.
The aryl group of the aryl group that may have a substituent found in the definition of R20, R21, R22 and R23 means phenyl group, 1-naphthyl group, 2-naphthyl group, anthracenyl group and the like.
The aryl of the arylalkyl group that may have a substituent found in the definition of R20, R21 and R22 has the same meaning as the above aryl group. The alkyl group in this case has the same meaning as the above lower alkyl group.
The heteroaryl group that may have a substituent found in the definition of R20, R21 and R22 means a group derived from a monocyclic ring or a condensed ring containing from 1 to 4 of at least one selected from the group consisting of a sulfur atom, an oxygen atom and a nitrogen atom. Examples thereof include pyrrolyl group, thienyl group, furyl group, thiazolyl group, oxazolyl group, isothiazolyl group, isoxazolyl group, imidazolyl group, pyrazolyl group, thiadiazolyl group, oxadiazolyl group, triazolyl group, tetrazolyl group, pyridyl group, a pyridazinyl group, a pyrimidinyl group, a pyrazinyl group, an indolyl group, an isoindolyl group, a benzothienyl group, a benzofuranyl group, an isobenzofuranyl group, a benzoimidazolyl group, an indazolyl group, a benzotriazolyl group, a benzothiazolyl group, a benzooxazolyl group, a quinolyl group, an isoquinolyl group, a cinnolinyl group, a phthalazyl group, a quinoxalyl group, a naphthyridyl group, a quinazolyl group, an imidazopyridyl group and the like.
The heteroarylakyl group that may have a substituent found in the definition of R20, R21 and R22 means those wherein the above heteroaryl is attached to any carbon atom of the above lower alkyl group.
The carbamoyl group that may have a substituent found in the definition of R1 and R2 means carbamoyl group having one or two substituents on the nitrogen atom thereof.
The substituent of the unsaturated heterocyclic ring that may be substituted with from 1 to 3 substituents, the aryl group that may have a substituent, the heteroaryl group that may have a substituent, the arylalkyl group that may have a substituent, the heteroarylalkyl group that may have a substituent, the heteroarylalkynyl group that may have a substituent, the arylalkoxy group that may have a substituent, the heteroarylalkoxy group that may have a substituent and the carbamoyl group that may have a substituent means hydroxyl group, a lower alkyl group, such as methyl, ethyl, n-propyl, isopropyl and the like, a lower alkoxy group, such as methoxy, ethoxy, n-propoxy, isopropoxy and the like, a halogen atom, such as a fluorine atom, a chlorine atom, a bromine atom, an iodine atom and the like, cyano group, an acyl group, such as anacetyl group, a propyonyl group, a benzoyl group and the like, amino group, nitro group, a carboxyl group that may have a protective group, carbamoyl group, an acylamino group, a sulfamoyl group, an alkylsulfonylamino group, an arylsulfonylamino group, a heteroaryl group, a carboxyalkyl group, a carboxyalkoxy group, a heteroarylalkyl group, a heteroarylalkoxy group, a methylenedioxy group, an ethylenedioxy group and the like, and selected from them.
The protective group of the carboxyl group that may have a protective group found in the definition of R1, R2, R20 and R21 means, for example, a lower alkyl group, such as methyl group, ethyl group, tert-butyl group and the like, a lower alkyl group substituted with a phenyl group that may have a substituent, such as p-methoxybenzyl, p-nitrobenzyl, 3,4-dimethoxybenzyl, diphenylmethyl, trityl, phenethyl and the like, a halogenated lower alkyl group, such as 2,2,2-trichloroethyl, 2-iodoethyl and the like, a lower alkanoyloxy lower alkyl group, such as pivaloyloxymethyl, acetoxymethyl, propyonyloxymethyl, butyryloxymethyl, valeryloxymethyl, 1-acetoxyethyl, 2-acetoxyethyl, 1-pivaloyloxyethyl, 2-pivaloyloxyethyl and the like, a higher alkanoyloxy lower alkyl group, such as palmitoyloxyethyl, heptadecanoyloxymethyl, 1-palmitoyloxyethyl and the like, a lower alkoxycarbonyloxy lower alkyl group, such as methoxycarbonyloxymethyl, 1-butoxycarbonyloxyethyl, 1-(isopropoxycabonyloxy)ethyl and the like, a carboxy lower alkyl group, such as carboxymethyl, 2-carboxyethyl and the like, a heteroaryl group, such as 3-phthalidyl and the like, a benzoyloxy lower alkyl group that may have a substituent, such as 4-glycyloxybenzoyloxymethyl and the like, a substituted dioxolene lower alkyl group, such as (5-methyl-2-oxo-1,3-dioxolene-4-yl)methyl and the like, a cycloalkyl-substituted lower alkanoyloxy lower alkyl group, such as 1-cyclohexylacetyloxyethyl and the like, and a cycloalkyloxycarbonyloxy lower alkyl group, such as 1-cyclohexyloxycarbonyloxyethyl and the like. Furthermore, it may be various acid amides. In its essence, any one can be the protective group of a carboxyl group as far as it can be a carboxylic acid by decomposition in vivo through some measures.
The ring in the term xe2x80x9cR20 and R21 may form a ring along with the nitrogen atom to which they are boundxe2x80x9d in the formula xe2x80x94NR20R21 found in the definition of Q means aziridine, azetidine, pyrrolidine, piperidine, perhydroazepine, perhydroazocine, piperazine, homopiperazine, morpholine, thiomorpholine, thiomorpholinedioxide, indoline, isoindoline, 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroisoquinoline, 2,3-dihydrobenzooxazine, 2,3-dihydrobenzothiadine, pyrrole, imidazole, pyrazole, triazole, tetrazole, indole, isoindole, indazole, benzotriazole and the like.
It is needless to say that the compounds having an asymmetric atom in the present invention, optical isomers thereof are also included in the invention. The present invention further includes the compounds of the present invention that are formed in vivo through metabolism.
The pharmacologically acceptable salt in the present invention includes, for example, an inorganic salt, such as a hydrochlorate, a hydrobromate, a sulfate, a phosphate and the like, an organic acid salt, such as an acetate, a maleate, a tartarate, a methanesulfonate, a benzenesulfonate, a toluenesulfonate and the like, and a salt formed with an amino acid, such as asparaginic acid, glutamic acid and the like.
The compounds of the present invention can be produced in such a manner that a heterocyclic ring condensed benzothiazine skeleton is produced by utilizing Smiles rearrangement reaction or Ullmann reaction of a heterocyclic ring-S-benzene ring compound obtained by a known method, and the xe2x80x94Dxe2x80x94Q side chain (in the formula, D and Q have the above meanings) is introduced to the NH group thus formed by a generally known method (for example, WO97/33871). Major general production reaction schemes of the compounds of the present invention will be shown below. 
In the formulae, the ring Het, R1 and R2 have the above meanings, and Hal represents chlorine, bromine or iodine. 
In the formulae, L represents a leaving group, M represents hydroxyl group that may have a leaving group or a protective group, and xxe2x80x2 represents an integer of 1 or 2. R represents a lower alkyl group, an aryl group that may have a substituent, a heteroaryl group that may have a substituent, an aryl lower alkyl group that may have a substituent or a heteroaryl lower alkyl group that may have a substituent. A and B are the same as or different from each other, and each represents hydrogen atom, a lower alkyl group, an aryl lower alkyl group that may have a substituent or a heteroaryl lower alkyl group that may have a substituent. The ring Het, R1 and R2 have the above meanings.
The benzothiazine compound (b or bxe2x80x2) obtained in Production Method 1 and a compound having at both ends thereof hydroxyl groups that may have a leaving group or both a leaving group and a protective group are reacted in the presence of a base, to obtain a benzothiazine compound represented by the formula (c). In this case, preferred examples of the base include potassium carbonate, sodium hydride, n-butyl lithium, t-butoxy potassium and the like. Preferred examples of the leaving group herein include halogen, sulfonate and the like. Examples of the protective group of the hydroxyl group include t-butyldimethylsilyl group, tetrahydropyranyl group and the like. As a reaction solvent, any one that does not participate in the reaction can be used. It is then treated with m-chloroperbenzoic acid (MCPBA) in methylene chloride, with monopersusfate (potassium peroxymonosulfate) in a mixed solvent of methanol and water, or with hydrogen peroxide in acetic acid, to obtain an oxide or a dioxide.
Next, in the case where M is hydroxyl group that may have a protective group, and for example in the case of hydroxyl group protected by a t-butyldimethylsilyl group, it is treated with tetrabutylammoniumfluoride in a solvent, such as tetrahydrofuran and the like, to form an alcohol (d), and then treated with a hydrazine hydrate to obtain an amine (e). In the case where M is a leaving group, the amine can also be obtained in the same manner by treating with phthalimide potassium. The amine (e) and an aldehyde compound are subjected to dehydration condensation to form Schiff base, which is treated with a reducing agent, such as sodium boron hydride and the like, to obtain an amine compound represented by the formula (f). As the reaction solvent, any one that does not participate in the reaction can be used.
Furthermore, in the case where M is a leaving group, a secondary or tertiary amine compound (g) can be obtained by acting a primary or secondary amine.
Resolution of optical isomers is conducted by chiral column chromatography or by fractional recrystallization of a salt with an optically active organic acid or organic amine.
Next, to describe the usefulness of the present invention, Pharmacological Experimental Examples will be shown below.
(1) Inhibitory Effects on Various Mediators Release from Rat Basophilic Leukemia Cell Line (RBL-2H3)
RBL-2H3cells (i.e., a cell line originating in rat cells), by stimulation with IgE sepecific antigen, not only release histamine and serotonin but also release and produce cytokines such as TNF xcex1 and prostaglandins which are inflammatory mediators. In this experimental system, inhibitory effects on various mediators release were examined by using serotonin as an indication.
The cells were beforehand labeled with [3H]-labeled serotonin and, at the same time, sensitized with the IgE antibody. After incubating with the compounds of the present invention, they were stimulated with the specific antigen. Then the inhibitory activity of each compound was calculated from the amount of the [3H]-labeled serotonin thus liberated into the medium due to the stimulation and the amount of [3H]-labeled serotonin liberated when no compound of the present invention was added.
The results are shown in Table 1.
The compound numbers in the table correspond to Example Nos. as will be given hereinafter (the same will apply hereinafter).
(2) Inhibitory Effects on Various Mediators Release from Human Basophils
6 ml of 6% dextran (for separating leukocytes, having a high molecular weight) was added to 20 ml of heparinized blood. After stirring well, the resulting mixture was allowed to stand at 37xc2x0 C. for 30 min and thus erythrocytes were precipitated. The upper layer was taken up and phosphate buffered saline was added thereto followed by centrifugation at 185 g for 8 minutes to give a crude leukocyte fraction. These cells were subjected to hypotonic hematolysis and then suspended in D-PBS(+) containing 0.1%-BSA. The resulting suspension was used in the subsequent experiment as the leukocyte fraction containing basophils. 0.4 ml of this cell suspension was preliminarily heated to 37xc2x0 C. for 5 min and then 0.05 ml of a specimen solution was added thereto followed by a pretreatment at 37xc2x0 C. for 15 min. Next, 0.05 ml of a mite antigen solution was added thereto to induce an antigen-antibody reaction. After 10 min, the reaction was ceased by ice-cooling. Then the reaction mixture was centrifuged again at 185 g for 10 min and histamine and peptide leukotrienes in the resulting supernatant were determined by using enzyme immunoassay kits. From the results of the assay, the activities of the a heterocyclic ring condensed benzothiazine compound of inhibiting the liberation of histamine and peptide leukotriene were determined.
The results are given in Table 2, wherein the term xe2x80x9cleukotrienexe2x80x9d means peptide leukotriene.
(3) Production of Passive Sensitization Rat Allergic Air Pouch Type Inflammation Model and Measurement of Action of Compound
Rats (Sprague-Dawley series, male, 6 weeks) were anaesthetized with ether, and 6 ml of air is injected subcutaneously on a region of back to produce a circular or ovoid sape air pouch. 1 ml of mouse monoclonal anti-2,4-dinitrophenyl (DNP) IgE antibody was injected into the air pouch to subject the rats to passive sensitization. On the following day, the rats were anaesthetized with ether, and 0.9% physiological saline containing 1% Evans Blue dye and 1 mg/ml of DNP bovine serum albumin (DNP-BSA) was administered intravenously at a dose of 0.2 ml per 100 g body weight to initiate an allergic reaction. After 5 minutes, the rats were anaesthetized with ether, and 5 ml of 0.9% physiological saline containing 0.1% of bovine serum albumin was injected in the air pouch. After lightly kneading the air pouch, 2 ml of the physiological saline was collected. The amounts of histamine and peptide leukotriene in the collected physiological saline were measured by ELISA method.
The compound of the present invention was orally administered 30 minutes before the administration of DNP-BSA as an antigen, to examine the inhibitory action on the model.
The results are shown in Table 3.
Thus, the compounds of the present invention inhibit liberation of a chemical mediators, such as serotonin, histamine, leukotriene and the like, and the effect has been also confirmed in the inflammation model. Therefore, the compounds of the present invention can be used as a preventive or therapeutic agent for disease caused by liberation of a chemical mediators, such as serotonin, histamine, leukotriene and the like. More specifically, it is useful for prevention and remedy of allergic disease such as asthma, allergic coryza, atopic dermatitis, hives, hay fever, gastrointestinal allergy, food allergy and the like.
Moreover, the compounds of the present invention are useful from the viewpoint the low toxicity and high safety thereof.
When the compounds of the present invention are used for the above-mentioned diseases, they may be administered both orally and parenterally, and in the dosage form of tablets, powders, granules, capsules, syrups, troches, inhalants, suppositories, injections, ointments, ophthalmic ointments, eye drops, nasal drops, ear drops, cataplasmas, lotions, etc.
The administration dose widely varies depending on the type of the disease, the severity of the symptoms, the age, sex and drug sensitivity of the patient. In general, such a compound is administered in a daily dose of from about 0.03 to 1,000 mg, preferably from 0.1 to 500 mg and still preferably from 0.1 to 100 mg once to several times a day. In the case of injections, the dose usually ranges from about 1 xcexcg/kg to 3,000 xcexcg/kg, preferably from about 3 xcexcg/kg to 1,000 xcexcg/kg.
The compounds of the present invention may be processed into preparations by conventional methods with the use of conventional pharmaceutical carriers.
Namely, solid preparations for oral administration are prepared by mixing the principal agent with fillers, binders, disintegrating agents, lubricants, coloring agents, corrigents, antioxidants, etc. and then processed into tablets, coated tablets, granules, powders, capsules, etc. by conventional methods.
Examples of the above-mentioned fillers are lactose, corn starch, sucrose, glucose, sorbitol, crystalline cellulose, silicon dioxide, etc.
Examples of the binders are polyvinyl alcohol, polyvinyl ether, ethylcellulose, methylcellulose, acacia, tragacanth, gelatin, shellac, hydroxypropylcellulose, hydroxypropylmethylcellulose, calcium citrate, dextrin and pectin. Examples of the lubricants are magnesium stearate, talc, polyethylene glycol, silica, hardened vegetable oils, etc.
The coloring agents are those admitted to be added to medicines. Examples of the corrigents include cocoa powder, menthol, aromatic powder, peppermint oil, borneol and powdered cinnamon bark. As the antioxidants, use can be made of any pharmaceutically authorized ones such as ascorbic acid and xcex1-tocopherol. Needless to say, tablets and granules may be appropriately coated with sugar, gelatin, etc., if necessary.
Meanwhile, injections, eye drops, etc. can be prepared by blending the principal agent with, if needed, pH regulating agents, buffer agents, suspending agents, dissolution aids, stabilizers, tonicity agents, antioxidants, preservatives, etc. and then processed in a conventional manner. In such a case, it is also possible, if needed, to give freeze-dried preparations. Injections may be intravenously, hypodermically or intramuscularly administered.
Examples of the above-mentioned suspending agents include methylcellulose, Polysorbate 80, hydroxyethylcellulose, acacia, tragacanth, sodium carboxymethylcellulose and polyoxyethyelne sorbitan monolaurate.
Examples of the dissolution aids are polyoxyethylene-hardened castor oil, Polysorbate 80, nicotinamide, polyoxyethylene sorbitan monolaurate, etc.
Examples of the stabilizers usable herein include sodium sulfite, sodium metasulfite and ether. Examples of the preservatives usable herein include methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, sorbic acid, phenol, cresol and chlorocresol.
Ointments can be produced by blending the principal agent with, if needed, stabilizers, antioxidants, preservatives, etc. and processed in a conventional manner.